Localization of oxytocin receptor mRNA in the ovine uterus during the oestrous cycle and early pregnancy

ABSTRACT A synthetic 45-mer oligonucleotide corresponding to part of the ovine endometrial oxytocin receptor cDNA was hybridized to sections of ovine uterus collected from 40 ewes at different stages during the oestrous cycle, the first 3 weeks of pregnancy and seasonal anoestrus. The quantity of oxytocin receptor mRNA was measured as the optical density (OD) value on autoradiographs using image analysis. Message first appeared in the luminal epithelium on days 14–15 of the cycle, increasing to a peak OD of 0·48 at oestrus and decreasing again between days 2 and 5. Oxytocin receptor mRNA in the superficial glands, deep glands and caruncular stroma increased between day 15 and oestrus to peak OD values of 0·17, 0·11 and 0·11 respectively, declining again by day 2 and reaching basal values (OD<0·015) by day 5. Hybridization to the myometrium tended to rise from a mean OD value of 0·01 on days 2–15 to a peak of 0·03±0·01 (mean±s.e.m.) on days 0–1, but the change was not significant. In pregnant ewes there was no detectable oxytocin receptor mRNA on days 14–15 in any region, but hybridization to the luminal epithelium was present in two of three ewes on day 21. In anoestrous ewes oxytocin receptor mRNA concentrations in all areas of the endometrium were approximately half those measured at oestrus. Optical density readings for oxytocin receptor mRNA in the various uterine compartments were compared with measurements of oxytocin receptors in the same regions as assessed by binding studies using the 125I-labelled oxytocin antagonist d(CH2)5[Tyr(Me)2,Thr4,Tyr-NH29]-vasotocin (125I-labelled OTA). In the endometrium, receptor mRNA and 125I-labelled OTA binding patterns changed in parallel, and both sets of measurements were significantly correlated (P<0·01). In the myometrium, a significant increase in 125I-labelled OTA binding occurred at oestrus; this was not accompanied by a similar increase in oxytocin receptor mRNA hybridization. This study helps to confirm that the previously identified cDNA clone is derived from the ovine oxytocin receptor, as patterns of oxytocin receptor mRNA expression in the endometrium closely resembled those of oxytocin binding. Maximum expression and binding both occurred at oestrus, suggesting that regulation of the oxytocin receptor gene in the uterus occurs principally at the transcriptional, rather than at the translational, level. Failure to detect a significant increase in myometrial mRNA expression at oestrus may indicate that the endometrial and myometrial oxytocin receptors are of different isoforms.

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Regulation of oxytocin receptor gene expression in sheep: tissue specificity, multiple transcripts and mRNA editing

Reproduction ◽

10.1530/reprod/120.1.187 ◽

2000 ◽

pp. 187-200 ◽

Cited By ~ 2

Author(s):

HC Feng ◽

M Bhave ◽

RJ Fairclough

Keyword(s):

Gene Expression ◽

Mrna Expression ◽

Receptor Gene ◽

Oxytocin Receptor ◽

Oestrous Cycle ◽

Receptor Expression ◽

Mrna Editing ◽

Oxytocin Receptor Gene ◽

Receptor Mrna ◽

Receptor Gene Expression

The increase in uterine oxytocin receptor concentrations over the late luteal phase of the oestrous cycle in sheep is thought to play an important role in the regulation of the duration of the cycle by facilitating the effect of oxytocin on uterine prostaglandin release. Experiments indicated that oxytocin receptor mRNA expression in the endometrium was high at oestrus compared with at days 2, 7 and 12 of the oestrous cycle. The amount of oxytocin receptor mRNA expression in the pituitary gland did not show any significant differences during the oestrous cycle. Oxytocin receptor cDNA was obtained and characterized from ovine uterine endometrium on day 15 of the oestrous cycle, using RT-PCR techniques, to study the mechanisms underlying the resolution of oxytocin receptor expression. The cDNA sequence for the oxytocin receptor gene in sheep was found to be similar to that described previously, except for a difference of seven nucleotides. These nucleotide differences resulted in changes in four of the deduced amino acids in the oxytocin receptor sequence. The heterogeneity of the different sized oxytocin receptor transcripts in sheep is due, at least in part, to the alternative use of polyadenylation sites. Northern hybridization confirmed that the oxytocin receptor gene is expressed in ovine corpus luteum. The investigations on oxytocin receptor gene expression indicate that the patten of oxytocin receptor gene expression in sheep is not only tissue-specific, but also highly function-related. Evidence was obtained of mRNA editing in both the coding and the 3'-untranslated (3'UTR) regions of oxytocin receptor gene transcripts in ovine endometrium; this was the first demonstration of this phenomenon for oxytocin receptor mRNA. The present results indicate that the observed differences in oxytocin receptor mRNA sequences for the different oxytocin receptor populations in endometrium are due to mRNA editing. mRNA editing of oxytocin receptor transcripts may be reflected in changes in the amino acid composition of the carboxyl terminus of the receptor, which would explain the differences in the observed responses to an oxytocin challenge.

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Autoradiographic localization of oxytocin receptors in the endometrium during the oestrous cycle of the ewe

Journal of Endocrinology ◽

10.1677/joe.0.1300199 ◽

1991 ◽

Vol 130 (2) ◽

pp. 199-NP ◽

Cited By ~ 11

Author(s):

V. J. Ayad ◽

E. L. Matthews ◽

D. C. Wathes ◽

T. J. Parkinson ◽

M. L. Wild

Keyword(s):

Epithelial Cells ◽

Stromal Cells ◽

Oxytocin Receptor ◽

Oestrous Cycle ◽

Luminal Epithelium ◽

Tissue Samples ◽

Luteal Regression ◽

Oxytocin Receptors ◽

Secretory Glands ◽

Luminal Epithelial Cells

ABSTRACT The present study was designed to determine the localization of the endometrial oxytocin receptor during the ovine oestrous cycle, particularly on day 14, the time of initiation of luteal regression in the ewe. Samples were obtained from 29 ewes at different stages of the oestrous cycle (several during the luteal phase and on every day between day 14 (− 2) and day + 3 of the oestrous period). Oxytocin receptors were localized autoradiographically in sections of uterine tissue, using the 125I-labelled oxytocin receptor antagonist [1-(β-mercapto-β,β-cyclopentamethylene propionic acid), 2-(ortho-methyl)-Tyr2,Thr4,Orn8,Tyr9-NH2]-vasotocin (125I-labelled OTA). There was some variation in the pattern of 125I-labelled OTA labelling between different uterine tissue samples from the same ewe and also between samples obtained from different ewes thought to be at the same stage of the oestrous cycle. A clear overall pattern did, however, emerge with 125I-labelled OTA-binding sites distributed between luminal epithelial cells, glandular epithelial cells and caruncular stromal cells to varying extents on different days of the cycle. During the luteal phase (days 5–12) clear specific labelling of endometrial tissue was generally absent. On day 14 labelling was evident on the luminal epithelium, but only in nine tissue samples out of a total of 18 studied, indicating that the entire luminal surface did not contain oxytocin receptors at this time. Between the day before oestrus and day 3 of the oestrous cycle the luminal epithelium was consistently labelled. The most extensive labelling of the remaining endometrial tissue was observed on the day of oestrus, with 125I-labelled OTA-binding sites clearly present on the stromal cells within caruncles and on a large proportion of secretory epithelia. This contrasted with the day before and the day after oestrus when labelling of glandular tissue was confined to the superficial endometrium, and labelling of caruncular stromal cells, although sometimes evident, was never as intense as on day 0. On days 2 and 3 labelling varied between being similar to that found on day 1 and being confined to the luminal epithelium and very few superficial secretory glands. The results of this study lead us to conclude that the oxytocin receptor shows a differential distribution between stromal cells, epithelial cells lining secretory glands and luminal epithelial cells during the oestrous cycle; that the steroidal regulation of the oxytocin receptor differs between endometrial cell types; and that control of the luminal epithelial oxytocin receptors is probably of particular importance to the regulation of prostaglandin F2α release at luteal regression and during the maternal recognition of pregnancy. Journal of Endocrinology (1991) 130, 199–206

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Regulation of oxytocin, oestradiol and progesterone receptor concentrations in different uterine regions by oestradiol, progesterone and oxytocin in ovariectomized ewes

Journal of Endocrinology ◽

10.1677/joe.0.1510375 ◽

1996 ◽

Vol 151 (3) ◽

pp. 375-393 ◽

Cited By ~ 62

Author(s):

D C Wathes ◽

G E Mann ◽

J H Payne ◽

P R Riley ◽

K R Stevenson ◽

...

Keyword(s):

Epithelial Cells ◽

Low Dose ◽

Progesterone Receptors ◽

Cell Types ◽

Oxytocin Receptor ◽

Luminal Epithelium ◽

Time Points ◽

Progesterone Treatment ◽

Oxytocin Receptors ◽

Receptor Concentration

Abstract The regulation of oxytocin, oestradiol and progesterone receptors in different uterine cell types was studied in ovariectomized ewes. Animals were pretreated with a progestogen sponge for 10 days followed by 2 days of high-dose oestradiol to simulate oestrus. They then received either low-dose oestradiol (Group E), low-dose oestradiol plus progesterone (Group P) or low-dose oestradiol, progesterone and oxytocin (via osmotic minipump; Group OT). Animals (three to six per time-point) were killed following ovariectomy (Group OVX), at oestrus (Group O) or following 8, 10, 12 or 14 days of E, P or OT treatment. In a final group, oxytocin was withdrawn on day 12 and ewes were killed on day 14 (Group OTW). Oxytocin receptor concentrations and localization in the endometrium and myometrium were measured by radioreceptor assay, in situ hybridization and autoradiography with the iodinated oxytocin receptor antagonist d(CH2)5[Tyr(Me)2,Thr4,Tyr-NH29]-vasotocin. Oestradiol and progesterone receptors were localized by immunocytochemistry. Oxytocin receptors were present in the luminal epithelium and superficial glands of ovariectomized ewes. In Group O, endometrial oxytocin receptor concentrations were high (1346 ± 379 fmol [3H]oxytocin bound mg protein−1) and receptors were also located in the deep glands and caruncular stroma in a pattern resembling that found at natural oestrus. Continuing low-dose oestradiol was unable to sustain high endometrial oxytocin receptor concentrations with values decreasing significantly to 140 ± 20 fmol mg protein−1 (P<0·01), localized to the luminal epithelium and caruncular stroma but not the glands. Progesterone treatment initially abolished all oxytocin receptors with none present on days 8 or 10. They reappeared in the luminal epithelium only between days 12 and 14 to give an overall concentration of 306 ± 50 fmol mg protein−1. Oxytocin treatment caused a small increase in oxytocin receptor concentration in the luminal epithelium on days 8 and 10 (20 ± 4 in Group P and 107 ± 35 fmol mg protein−1 in Group OT, P<0·01) but the rise on day 14 was not affected (267 ± 82 in Group OT and 411 ± 120 fmol mg protein−1 in Group OTW). In contrast, oestradiol treatment was able to sustain myometrial oxytocin receptors (635 ± 277 fmol mg protein−1 in Group O and 255 ± 36 in Group E) and there was no increase over time in Groups P, OT and OTW with values of 61 ± 18, 88 ± 53 and 114 ± 76 fmol mg protein−1 respectively (combined values for days 8–14). Oestradiol receptor concentrations were high in all uterine regions in Group O. This pattern and concentration was maintained in Group E. In all progesterone-treated ewes, oestradiol receptor concentrations were lower in all regions at all time-points. The only time-related change occurred in the luminal epithelium in which oestradiol receptors were undetectable on day 8 but developed by day 10 of progesterone treatment. Progesterone receptors were present at moderate concentrations in the deep glands, caruncular stroma, deep stroma and myometrium in Group O. Oestradiol increased progesterone receptors in the luminal epithelium, superficial glands, deep stroma and myometrium. Progesterone caused the loss of its own receptor from the luminal epithelium and superficial glands and decreased its receptor concentration in the deep stroma and myometrium at all time-points. There was a time-related loss of progesterone receptors from the deep glands of progesterone-treated ewes between days 8 and 14. These results show differences in the regulation of receptors between uterine regions. In particular, loss of the negative inhibition by progesterone on the oxytocin receptor by day 14 occurred only in the luminal epithelium, but is unlikely to be a direct effect of progesterone as no progesterone receptors were present on luminal epithelial cells between days 8 and 14. The presence of oxytocin receptors in the luminal epithelium of ovariectomized ewes suggests that oestradiol is not essential for oxytocin receptor synthesis at this site. Oestradiol was able to sustain its own receptor at all sites, but high circulating progesterone was always inhibitory to oestradiol receptors. In general, oestradiol stimulated progesterone receptors in epithelial cells whereas progesterone abolished its own receptor from epithelial cells over a period of time, but had a lesser effect on stromal cells. The concentration of all three receptors is therefore differentially regulated between different uterine cell types, suggesting the importance of paracrine effects which remain to be elucidated. Journal of Endocrinology (1996) 151, 375–393

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Oxytocin and oxytocin-receptor mRNA expression in the human gastrointestinal tract: a polymerase chain reaction study

Regulatory Peptides ◽

10.1016/j.regpep.2003.12.017 ◽

2004 ◽

Vol 119 (1-2) ◽

pp. 39-44 ◽

Cited By ~ 55

Author(s):

Hans-Jürg Monstein ◽

Niclas Grahn ◽

Mikael Truedsson ◽

Bodil Ohlsson

Keyword(s):

Polymerase Chain Reaction ◽

Gastrointestinal Tract ◽

Mrna Expression ◽

Oxytocin Receptor ◽

Human Gastrointestinal Tract ◽

Chain Reaction ◽

Reaction Study ◽

Receptor Mrna ◽

Polymerase Chain

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Oxytocin receptor mRNA expression in the ventromedial hypothalamus during the estrous cycle

Journal of Neuroscience ◽

10.1523/jneurosci.15-07-05058.1995 ◽

1995 ◽

Vol 15 (7) ◽

pp. 5058-5064 ◽

Cited By ~ 63

Author(s):

TL Bale ◽

DM Dorsa ◽

CA Johnston

Keyword(s):

Mrna Expression ◽

Estrous Cycle ◽

Ventromedial Hypothalamus ◽

Oxytocin Receptor ◽

Receptor Mrna

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The role of the endometrial oxytocin receptor in determining the length of the sterile oestrous cycle and ensuring maintenance of luteal function in early pregnancy in ruminants

Philosophical Transactions of the Royal Society B Biological Sciences ◽

10.1098/rstb.1994.0067 ◽

1994 ◽

Vol 344 (1309) ◽

pp. 291-304 ◽

Cited By ~ 22

Keyword(s):

Gene Expression ◽

Molecular Mechanisms ◽

Transmembrane Domain ◽

Receptor Gene ◽

Oxytocin Receptor ◽

Oestrous Cycle ◽

High Rate ◽

Endocrine Function ◽

Oxytocin Receptor Gene ◽

Receptor Gene Expression

The oxytocin receptor, a seven transmembrane domain, G protein-linked receptor molecule, plays a central role in determining the endocrine function of the ruminant uterine endometrium. During non- pregnant cycles the control of this molecule by circulating steroid hormones leads to regression of the corpora lutea. The kinetics of the mechanisms involved determine the time at which luteolysis occurs, and therefore the length of the oestrous cycle. In pregnancy, secretions of the trophoblast block endometrial oxytocin receptor gene expression and lead to luteal maintenance. An understanding of the molecular mechanisms involved in the steroidal control of oxytocin receptor gene expression will provide an explanation for the relative constancy of oestrous cycle lengths in non-pregnant animals. Unravelling the way in which trophoblast products block expression of the oxytocin receptor gene will lead to a better understanding of the reasons for the high rate of embryonic loss in domestic ruminants.

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Tissue dependent expression of bitter receptor TAS2R38 mRNA

10.1101/293399 ◽

2018 ◽

Author(s):

Jennifer E. Douglas ◽

Cailu Lin ◽

Corrine J. Mansfield ◽

Charles J. Arayata ◽

Beverly J. Cowart ◽

...

Keyword(s):

Mrna Expression ◽

Bitter Taste ◽

Receptor Gene ◽

Active Form ◽

Taste Receptor ◽

Human Populations ◽

Bitter Taste Receptor ◽

Receptor Mrna ◽

Inactive Form ◽

Autopsy Data

AbstractTAS2R38 is a human bitter receptor gene with a common but inactive allele and people homozygous for the inactive form cannot perceive certain bitter compounds. The frequency of the inactive and active form of this receptor is nearly equal in many human populations and heterozygotes (with one copy of the active form and another copy of the inactive form) have the most common diplotype. However, heterozygotes differ markedly in their perception of bitterness, in part perhaps because of differences in TAS2R38 mRNA expression. Other tissues including the nasal sinuses express this receptor, where it contributes to pathogen defense. We asked whether heterozygotes with high TAS2R38 mRNA expression in taste tissue were also likely to express more TAS2R38 mRNA in sinonasal tissue. To that end, we measured TAS2R38 bitter taste receptor mRNA by qPCR in biopsied tissue, and learned that expression amount of one is not related to the other. We confirmed the general independence of expression in other tissue expressing TAS2R38 mRNA using autopsy data from the GTEx project. Taste tissue TAS2R38 mRNA expression among heterozygotes is unlikely to predict expression in other tissues, perhaps reflecting tissue-dependent function and hence regulation of this protein.

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